chromosomal study of yellow sweet-clover (Melilotus officinalis (L.) Lam.) under heavy metals pollution of lead and zinc mine of Zeh Abad, Qazvin

Document Type : Research Paper


1 Department of Biology, Faculty of science, Payame-Noor University, Tehran, Iran

2 Department of Geology, Imam Khomeini International University, Quzvin, Iran


Melilotus officinalis (L.) Lam. belonging to Fabaceae growing in the vicinity of zinc and lead mine around Zeh abad village, Qazvin province, Iran. The aim of this study is determination of amount of zinc and lead pollution in sampled soil and water and find out how M. officinalis is affected by a long-term exposure to pollutants focusing on the meiotic behavior of pollen mother cells (PMCs) and pollen fertility. The chemical analysis of soil and water samples from the studied area by the ICP-MS indicates presence of lead and zinc in a concentration much higher than the standard. The results revealed that polluted soil and water with heavy metals had a significant impeding effect on the division of PMCs in M. officinalis as noticed by the increase in meiosis abnormality percentage, decrease in meiotic index (MI) and pollen fertility compared to the control and can lead to genotoxicity in under study plant. The present data indicate that this species is not meiotically stable. In general, the pollen mother cells of specimen grown in polluted soil exhibited an increased incidence of chromosome stickiness, B-chromosomes, chromosome bridges, laggard chromosomes, micronuclei, cytomixis and formation of tripolar cells.


Abubacker, M.N., Sathya, C., 2017. Genotoxic Effect of Heavy Metals Cr, Cu, Pb and Zn using Allium cepa L. Biosciences, Biotechnology Research Asia 14(3), 1181–1186.
Ackova, D.G., 2018. Heavy metals and their general toxicity on plants. Plant Science Today 5(1), 14–18.
Assistance of the human environment. Office of water and soil: Sewage outlet standard, 4 p.
Assistance of the human environment. Office of water and soil: Standards for quality of soil resources and its guides. 2014, 166 p.
Baptists-Giacomoelli, F.R., Pagliarini, M.S., Almeida, J.L., 2000. Elimination of micronuclei from microspores in a Brazilian oat (Avena sativa L.) variety. Genetics and Molecular Biology 23 (3), 681–684.
Bhuyan, S., Islam, S., 2017. A Critical Review of Heavy Metal Pollution and Its Effects in Bangladesh, Science Journal of Energy Engineering 5 (4), 95–108.
Bifeng, H., Xiaolin J., Jie, H., Dongyun, X., Fang, X., Yan, L., 2017. Assessment of Heavy Metal Pollution and Health Risks in the Soil-Plant-Human System in the Yangtze River Delta, China. International Journal of Environmental Research and Public Health 12, 1–18.
Biscupa A., Izmailow, R., 2004. Endosperm development in seed of Echium vulgare L. (Boraginaceae) from polluted sites. Acta Biologica Cracoviensia Series Botanica 46, 29–44.
Dalvand M., Hamidian A.H., Zare Chahooki M.A., Moteshare Zadeh B., Mirjalili S.A.A., Esmaeil Zade E., 2014. Comparing heavy metal accumulation abilities in Artemisia aucheri and Astragalus gummifer in Darreh Zereshk region, Taft, Desert 19(2), 137–140.
Dalvand M., Hamidian A.H., Zare Chahouki M.A., Mirjalili A.A., Motesharrezadeh B. Esmaeilzadeh E., 2016. Determination of the concentration of heavy metals (Cu, Pb, Zn) in roots of Artemisia sp. in natural lands of Darreh Zereshk copper mine, Taft, Yazd, Journal of Natural Environment 69 (1), 35–46.
Ghaffari, S.M., 2006. Occurrence of diploid and polyploidy microspores in Sorghum bicolor (Poaceae) is the result of cytomixis. African Journal of Biotechnolgy 5, 1450–1453.
Gomurge, A.N., 2000. Cytological effect of the herbicide 2,4-D isooctylester 48% on root mitosis of Allium cepa. Cytologia 65, 383–388.
Gottardini, E., Cristofolini, F., Paoletti, E., Lazzeri, P., Pepponi, G., 2004. Pollen viability for air pollution biomonitoring. Journal of Atmospheric Chemistry 49, 149–159.
Hamidian, A.H., Norouznia H., Mirzaei R., 2016. Phytoremediation efficiency of Nelumbo nucifera in removing heavy metals (Cu, Cr, Pb, As and Cd) from water of Anzali wetland, Journal of Natural Environment, 69(3), 669–679
Higashitani, A., 2013. High temperature injury and auxin biosynthesis in microsporogenesis. Frontier of Plant Sciences 4, 1–4.
Izmailow R., 2000. Reproduction of Viccia cracca L. in the polluted environment of the Legnica – Głogow copper basin (Poland). Acta Biologica Cracoviensia Series Botanica 42(2), 125–133.
Jones, N., Houben, A., 2003. B-chromosomes in plant: escapees from the A-chromosome genome? Trends in plant science 8(9), 417–423.
Klosowska, K., Izmailow, R., Muszynska, E., 2009. Wpływ warunkow skażonego siedliska poboczy szlakow komunikacyjnych na procesy embriologiczne u wybranych gatunkow roślin (Lotus corniculatus L. i Lepidium ruderale L.). Journal of Elementology 14, 47–48.
Koduru, P.R.K., Rao, M.K., 1981. Cytogenetics of synaptic mutants in higher plants. Theoretical and applied genetics 59(4), 197–214.
Koscinska-Pajak, M., 2000. Microspores and pollen grain in triploid Chondrilla juncea L. from unpolluted and polluted areas. Acta Biologica Cracoviensia Series Botanica 42(2), 135–140.
Kumar, G., Bhardwaj, M., 2017. Comparative genotoxicity of heavy metals in root meristems of Cuminum cyminum L. Chromosome Botany 12(3), 56–62.
Lattoo, S.K., Khan, S., Bamotra, S., Dhar, A.K., 2006. Cytomixis impairs meiosis and influences reproductive success in Chlorophytum comosum (Thunb) Jacq., an additional strategy and possible implications. Journal of Bioscience 31, 629–637.
Love, R.M., 1949. Estudos citológicos preliminares de trigos  rio-grandenses. Secretaria do Estado dos Negócios da Agricultura, Indústria e Comércio, Porto Alegre, 23p. (Circular 74).
Mansuelli, R.W., Tanimoto, E.Y., Brown, C., Comai, L., 1995. Irregular meiosis in a somatic hybrid between Solanum bulbocastanum and S. tuberosum detected by species-specific PCR markers and cytological analysis. Theoretical and applied genetics 91(3), 401–408.
Martinez-Sanchez M.J., Garcia-Lorenzo M.L., Perez- Sirvent C., Bech J., 2012. Trace element accumulation in plants from an aridic area affected by mining activities. Geochemical Exploration, 123, 8–12.
Micieta K., Murin G., 1996. Microspore analysis for genotoxicity of a polluted environment. Environmental and Experimental Botany 36(1), 21–27.
Nepi, M., Franchi, G.G., 2000. Cytochemistry of mature angiosperm pollen. Plant Systematics and Evolution 222, 45–62.
Nicklas, R.B., Ward, S.C., 1994. Elements of error correction in mitosis: microtubule capture, release and tension. Cell Biology 126(5), 1241–1253.
Nirmala, A., Rao, P.N., 1996. Genetics of chromosome numerical mosaism in higher plants. The nucleus 39, 151–175.
Norouznia H. Hamidian A.H. 2014. Phytoremediation efficiency of pondweed (Potamogeton crispus) in removing heavy metals (Cu, Cr, Pb, As and Cd) from water of Anzali wetland, International Journal of Aquatic Biology, 2(1): 206–214.
Ostrolucka M.G., 1989. Differentiation of male reproductive organs and oak fertility in urban environment. Biologia (Bratislava) 44(9), 793–799.
Pagliarini, M.S., 1990. Meiotic behavior and pollen fertility in Aptenia cordifolia (Aizoaceae). Caryologia 43(2), 157–162.
Pagliarini, M.S., 2000. Meiotic behavior of economically important plant species: the relationship between fertility and male sterility. Genetic and Molecular Biology 23, 997–1002.
Paupiere, M.J., Van Heusden, A.W., Bovy, A.G., 2014. The metabolic basis of pollen thermo-tolerance: Perspectives for Breeding. Metabolites 4, 889–920.
Rani, N., Kumar, K., 2016. Mutagenic Effects of EMS on Pollen Mother Cells of Catharanthus roseus (L.) G. Don. Chromosome Botany 11(3), 51–55.
Ranjbar, M., Hajmoradi, F., Karamian, R., 2012. An overview on cytogenetics of the genus Onobrychis (Fabaceae) with special reference to O. sect. Hymenobrychis from Iran. Caryologia. 65 (3), 187–198.
Ranjbar, M., Hajmoradi, Z., Karamian, R., 2011. Cytogenetic study and pollen viability of four populations of Trigonella spruneriana Boiss. (Fabaceae) in Iran. Journal of Cell and Molecular Research 3 (1), 19–24.
Ravindran, P.N., 1978. Cytological Irregularities Induced by Water Polluted with Factory Effluents: A preliminary report. Cytologia 43, 565–568.
Rechinger, K.H., 1982. Stachys. In: Rechinger, K.H. (Ed.), Flora Iranica, Graz: Akademische Druck- und Verlagsanstalt, 150, 354–396.
Rezanejad, F., 2013. The response of anther and pollen development, pollen cellular material release and pollen proteins to air pollution in Petunia hybrid Juss. (Solanaceae). Iranian Journal of Science and Technology, Transaction A: Science 1, 63–68.
Samardakiewicz, S., Wozny, A., 2005. Cell division in Lemna minor roots treated with lead, Aquatic Botany 83, 289–295.
Seregin, I.V., Ivanov, V.B., 2001. Physiological Aspects of Cadmium and Lead Toxic Effects on Higher Plants, Russian Journal of Plant Physiology 48 (4), 523–544.
Singhal, V.K., Kumar, P., 2008. Impact of cytomixis on meiosis, pollen viability and pollen size in wild populations of Himalayan poppy (Meconopsis aculeata Royle). Journal of Biosciences, 33: 371–380.
Slomka, A., Walec, P., Kellner, K., Jedrzejcyk-Korrycinsk, M., Rostanski, A., Kuta, E., 2010. Was reduced pollen viability in Viola Tricolor L. the result of heavy metal pollution or rather the tests applied? Acta Biologica Cracoviensia Series Botanica 52(1), 123–127.
Sotohian; F., Hojjati; L., Sharifi S., 2014. Environmental Effects of Zehabad-e-Qazvin Lead and Zinc Mine. J. Hum. Env. 28, 17–29.
Stace, C.A., 1991. Plant Taxonomy and Biosystematics. Cambridge University Press, Cambridge.
Tripathi, R., Girjesh, K., 2010. Genetic loss through heavy metal induced chromosomal stickiness in Grass pea. Caryologia 63 (3), 223–228.
Utsunomiya, K.S., Bione, N.C.P., Pagliarini, M.S., 2002. How many different kinds of meiotic abnormalities could be found in a unique endogamous maize plant? Cytologia 67(2), 169–176.
Vizintin, L., Bohanec, B., 2004. In vitro manipulation of Cucumber (Cucumis sativus L.) pollen and microspores: isolation procedures, viability tests, germination, maturation. Acta Biologica Cracoviensia Series Botanica 46, 177–183.
Wu, W., Wu, P., Yang, F., Sun, D.L., Zhang, D.X., Zhou, Y.K., 2018. Assessment of heavy metal pollution and human health risks in urban soils around an electronics manufacturing facility. Science of The Total Environment 630, 53–61.
Yucel, E., Hatdpoglu, A., Sozeni, E. Guner, S.T., 2008 The effects of the lead (PbCl2) on mitotic cell division of Anatolian Black Pine (Pinus nigra ssp. pallasiana) Biological Diversity and Conservation 1 (2), 124–129.
Zohari, S., Khatooni, S. Zaidi, S., 2012. Cytological Studies on 14 plant species under polluted conditions. Pakestan Journal of Botany 44(6), 1977–1982.